Use of bone marrow somatic cell hybrid lines to generate monoclonal antibodies specifically reactive with rare marrow cells.
The technique of somatic cell hybridization has been applied to obtain new sources of immunogen for monoclonal antibody production. A marrow population enriched for rare and undifferentiated cells was hybridized with A9 cells. Mature cells were depleted by using mice at 8 days following 5-fluorouracil treatment and by using a sorting procedure. Selection of hybrids was in medium containing hypoxanthine, aminopterin, and thymidine (HAT). Chromosome analysis was used to verify that clones contained hybrid cells. An example of an antibody (H513E3) obtained by immunizing with a hybrid cell line (H5Scl1.4.4) is presented. The H513E3 antibody showed low reactivity with normal marrow (0%-3%), and hemopoietic cell lines of various types exhibited no cross-reaction. However, about 10% of cells from the adherent stromal layer of long-term marrow cultures reacted with the H513E3 antibody. A specific cell type was labeled that appeared to have endothelial-like morphology when observed with immunogold labeling and electron microscopy. The hybridization technique allows a cell of a particular differentiation lineage to be conserved, wholly or partially, in a cloned form, thus overcoming the heterogeneity of a normal marrow population.
Hasthorpe
S
S
Cell Biology Unit, Peter MacCallum Cancer Institute, Melbourne, Australia.
Rogerson
J
J
Green
S L
SL
Radley
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D005260:Female; D005347:Fibroblasts; D005434:Flow Cytometry; D005472:Fluorouracil / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006822:Hybrid Cells / Q000276:immunology*; D007114:Immunization; D007621:Karyotyping; D008264:Macrophages / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008854:Microscopy, Electron
D000911:Antibodies, Monoclonal; D005472:Fluorouracil